Urinalysis

Jorge Anton D. Ordas

Eloisah Vin S. Ragodon
Lindley C. Susi
Mercer Keith Von Possel
Mark Rainier C. Yu
3BIO6

Urine

Is a composite of various organic and

inorganic substances which are normally
waste products from metabolic processes.

Body fluids filtered by the kidney and

excreted through the urethra

Typically yellow-amber in color

Used in testing for physiological

imbalances or disease

Urine (composition)

Average Urine
Output(Adults): 1-2
liters per 24 hours

5% - solutes
95% - water
Main Organic
Constituents: Urea,

Creatinine, Uric acids,

Ammonium, SO4 /Sulfate,
and PO4.

Main Ions present:

Sodium, Potassium, Chloride,

Magnesium and Calcium ions

URINE (Output/Volume)

Oliguria
Less

than 400 ml of urine in 24 hours

Anuria
Less

than 100 ml of urine in 24 hours

Possible causes:
1.Prerenal

dehydration, congestive heart failure and

hemorrhage
2.Postrenal obstruction of the urinary tract
3.Renal parenchymal disease - acute tubular necrosis,
chronic renal failure

Urine (increase)

Polyuria
More

than 2L of urine in 24 hours

Possible causes:
1.Physiological states:

increased water intake/

excessive hydration, some drugs/ diuretic effects, intravenous
solutions
2.Pathologic
insipidus

states:

Diabetes mellitus, Diabetes

Urinalysis

Diagnostic tool used for the overall

evaluation of renal function or possible
malfunction/disease
Rapid detection and measurement of various
compounds that pass through the urine
Includes 2 steps of examination:
Physical

Characterization color, turbidity, presence of

suspended particles
Chemical Reaction Tests detection of normal and
pathologic organic constituents

Urinalysis (Importance)

To identify or detect the presence if excess

protein, glucose, bile pigments are left
unfiltered by the renal tubules, intact or
hemolysed RBCs and elevated level of
intestinal microbiota and possible etiologic
agent of urinary tract infection and renal
diseases

Objectives

To make an initial examination of the

urine sample

To examine the normal organic

constituents of the urine sample

To examine for pathologic organic

constituents of the urine sample by
comparing with positive control set-ups

Procedure/Methodology
A. Initial Examination of Urine
Note time of collection
Indicate the color and the turbidity and
check for suspended materials
Check pH through litmus paper

Procedure/Methodology
B. Examination for Normal Organic
Constituents
B1. Test for urea

1 mL urine
sample
-add

0.5 mL 70%
NaOH and 4 drops
bromine water
Observe
formation of
bubbles

Procedure/Methodology
B2. Test for uric acid
1 mL Urine
Sample

-add 5 mL of 20% Na2CO3

, then mix
-add 5 drops
phosphotungstic acid
reagent,
mix
-observethen
for formation
of blue solution
Record
Result

Procedure/Methodology
B.3 Indican test
5 5mL
mLUrine
Urine
Sample
Sample

- add 5 mL Obermayers
Reagent, then mix well
- add 3 mL chloroform.
Shake and allow
chloroform
to formation
settle.
- observe for
of blue color in lower
layer

Record Result
Result
Record

Procedure
B4. Creatinine test
2 mL Urine
Sample

-add 1 mL of alkaline
picrate solution
-note for formation of an
orange solution
Record Result

Procedure
C. Examination of pathologic organic
constituents
Controltest
C1.Positive
Gunnings
5 mL Urine
5 mL Urine + 2 mL
acetone

Sample
-Basify with 5 drops
concentrated ammonium
hydroxide .Check pH by red
litmus
paper. solution to
- add Lugols
sample until black cloud
-appears
note the formation of
iodoform crystals

Record Result

Procedure
C.2. Benedicts test
Positive Control

1 mL Urine + 1 mL
glucose solution

0.5 mL Urine
Sample

- add 5 mL of Benedicts
reagent
- heat in boiling water
bath for 2-3 minutes
then
- notecool
precipitate
formation
Record Result

Procedure
C3. Extons test
Positive Control

3 mL Urine
Sample

3 mL Urine + pinch
of albumin

-add 3 mL of Extons
reagent
-warm if cloudiness
appears
-if cloudiness persist or
increase in heating,
albumin is present
Record Result

Procedure
C4.Positive
Smiths
test
Control
3 mL Urine + 3-5
drops bile
pigment

5 mL Urine
Sample

-incline test tube and

overlay with 3 mL of
tincture of alcoholic
iodine mixture

Note if emerald
green is seen at
point of contact

Procedure
C5. Test for occult blood
Positive Control

3 mL acidified
Urine + 3 drops
of blood

3 mL Acidified
Urine Sample
- add 5 mL of 95% ethanol to half
spatula guaiac powder then add 5
mL of hydrogen peroxide in one
test
- addtube
5 mL of this solution to 3
mL acidified urine and positive
- note the formation of iodoform
conrol
crystals

Record Result

Results

A. Initial Examination of Urine

Sample
Time
Color
Turbidity

pH

collected
9/20/12 7:15
am

Dark yellow

Turbid

9/20/12 7:30
am

Yellow Orange

Clear

9/20/12 9:00
am

Light Pale
Yellow

Slightly Turbid

9/19/12 10:59
pm

Light Yellow

Slightly turbid
w/ minute
suspended
particles

9/20/12 6:00
am

Dark Yellow

Clear

9/19/12 8:25
pm

Dark Yellow

Clear

9/20/12 7:45
am

Light Yellow

Clear

9/20/12 10:00
am

Yellow Orange

Turbid

9/20/12 9:55
am

Very light
yellow

Clear

10

9/20/12 9:20

Light Yellow

Clear

B. Examination for Normal Organic Constituents of

Urine Group
Sample
Test for Urea
1

White gas was observed then brown gas almost

immediately

Clear yellow solution upper layer and clear light yellow

lower layer

Light yellow solution, with N2

gases, presence of cloudy white substance

Formation of bubbles, light yellow solution

Formation of bubbles on top

Clear yellow solution with formation of bubbles on the

uppermost portion

Clear yellow solution with presence of bubbles and foamy

layer

Light yellow color upper layer with white fumes and clear
yellow second layer

Yellow solution with presence of bubbles indicating

formation of N2 gas

10

Evolution of gas was observed

11

There was an evolution of N2

Group

Test for Uric Acid

Clear blue solution

Clear light blue solution

light blue colored solution appeared

Clear blue solution

Formation of blue clear solution

Clear blue solution

Clear light blue solution

Blue solution formed

Light blue solution

10

Light blue colored solution

11

Clear blue solution

Group

Indican Test

Clear blue colored lower layer and dark green upper layer

Clear brownish green upper layer and blue lower layer with
presence of emulsion

Olive green solution at upper layer and pale clear blue

solution at lower layer

Lower light blue clear solution, middle olive green clear

solution, upper thin black layer

Olive green upper layer and clear light blue lower layer

Clear brown upper layer and very light clear blue lower
layer

Green colored solution upper layer with clear blue lower

layer

No blue color formed at lower layer

Light blue solution at lower layer

10

Light blue colored solution at lower layer

11

Presence of the blue color in the lower layer

Group

Creatinine Test

Clear orange colored solution formed

Clear red orange solution

Dark orange colored solution appear

Red orange colored solution formed

Clear red orange solution formed

Clear red orange solution formed

Clear red orange solution formed

Orange solution formed

Orange solution formed

10

Red colored solution

11

Red orange colored solution

C. Examination for Pathological Constituents of Urine

Sample
C1. Gunnings Test
Group

Urine Sample

Positive Control

Black precipitate settled at

lower layer and cloudy layers
visible on upper layer

Black precipitate was formed

but disappeared. Turbid
solution with cloudy
precipitates at bottom

Dark yellow orange solution

with light yellow precipitate

Dark yellow solution with light

yellow precipitate

Light yellow turbid solution with

black cloud formation seen at
the lower layer/bottom

Pale turbid yellowish solution

with light yellow precipitate
at the bottom; no black cloud
solution appeared

Clear light yellow solution with

cloudy precipitate

Clear yellow solution with

yellow precipitate

Formation of cloudy white

yellow solution

Formation of white cloudy

precipitate with light yellow
orange solution

Group

Urine Sample

Positive Control

Cloudy yellow solution with

yellow suspension

Clear yellow orange solution

with cloudy white suspension
at the lower part

Turbid/cloudy solution with

precipitate on middle

Slightly turbid/cloudy yellow

solution with black precipitate
at bottom

Clear red orange solution with

cloudy white lower layer and
red orange precipitate at the
bottom

Clear orange solution with

cloudy white lower layer and
yellow orange precipitate at
bottom

Presence of black crystals

settled on the bottom

Presence of black crystals

settled on the bottom

10

Formation of black iodoform

crystals

Turbid solution

11

Yellow solution with cloudy red

precipitate on the bottom

Yellow solution with white

precipitate on the bottom

C2. Benedicts Test

Group

Urine Sample

Positive Control

Blue Solution

Yellow-green turbid solution

with yellow precipitate settled
on the bottom

Semi turbid solution with green

precipitate

Turbid orange solution with

orange precipitate

Blue-green colored solution

appeared with formation of a
cloudy-like substance at the
lower part of the test tube

Two layers appeared: a dark

yellow turbid upper layer
solution and a greenish
colored solution for the lower
layer; yellow powdered-like
precipitate formed

Clear blue solution

Upper yellow layer, middle

green layer with orange
precipitate

Pale orange precipitate with

blue-green interphase; yellow
solution

Formation of a clear aqua

solution with suspended
particles

Group

Urine Sample

Positive Control

Clear green upper layer and

clear light blue lower layer with
some particles suspended in
the lower layer and between 2
layers

Orange solution with green

and orange precipitate

Orange solution upper layer;

Green turbid solution middle
layer; Brownish orange
precipitate at lower layer

Slightly clear blue solution

with little precipitate

Blue green turbid upper layer

with few scattered and settling
white precipitate

Turbid yellowish solution

through-out 4 layers with the
third layer being blue green;
Greenish and yellow
precipitate at the bottom

Clear blue solution

Presence of brick red

precipitate

10

Light blue solution

Orange precipitate formed in

an orange solution
Green turbid solution on top

C3. Extons Test

Group

Urine Sample

Positive Control

Clear light yellow solution

Creamy white solution with white

precipitate

Turbid light orange solution with

fibrous materials in upper layer and
turbid green with precipitate at lower
later

Clear orange solution

Clear light yellow solution

Cloudy pale yellow solution and

formation of light yellow
precipitate at the upper surface

Clear solution

Formation of cloudy mass

suspended in a solution

Formation of clear light yellow

solution

Formation of cloudy solution with

clear bottom

Clear light yellow-orange solution

Light yellow solution with light

yellow aggregates

Clear light yellow solution

White turbid solution with white

precipitate

Clear golden yellow solution

White turbid solution

Clear solution

Cloudiness is seen with mass of

white precipitate

10

Clear colorless solution

White cloudy present

C4. Smiths Test

Group

Urine Sample

Positive Control

Green interphase

Green interphase

No emerald green interphase

Emerald green interphase

Emerald green interphase

Emerald green interphase

Yellow interphase

Yellow interphase

No emerald green interphase

No emerald green interphase

No emerald green interphase

No emerald green interphase

No emerald green interphase

No emerald green interphase

No emerald green interphase

No emerald green interpahse

No emerald green interphase

seen at point of contact

No emerald green interphase at

point of contact

10

No emerald green interphase

Emerald green color appeared

at point of contact

11

Emerald green color is observed

at the point of contact

Emerald green color is

observed at the point of
contact

C5. Test for Occult Blood

Group

Urine Sample

Positive Control

Blue green ring absent

Blue green ring formed

Blue green ring absent

Blue green ring absent

Dark blue colored solution

appeared

Blue green ring absent

Blue green ring formed

Blue green ring absent

Blue green ring formed

Blue green ring absent

Blue green ring formed

Blue green ring absent

Blue green ring formed

Blue green ring absent

Blue green ring formed

Blue green ring absent

Blue green ring formed

10

Blue green ring absent

Blue green ring formed

11

Blue green ring absent

Blue green ring formed

Blue green ring formed

Discussion
a. Initial examination of urine sample

A. Initial Examination of Urine

Sample
A1. Time of collection

First voided morning specimen (most common)

Random urine sample (emergency)

Most ideal to test for substances

Most concentrated
Formed elements are more stable

Collected at any time, usually daytime

Exercise and excessive fluid intake can directly affect urine
composition

Clean-catch, midstream specimen (urine

culture)

Prior hydration of patient will lead to a clean catch urine

specimen

Time
collected

Color

Turbidity

pH

9/20/12 7:15
am

Dark yellow

Turbid

9/20/12 7:30
am

Yellow Orange

Clear

9/20/12 9:00
am

Light Pale
Yellow

Slightly Turbid

9/19/12 10:59
pm

Light Yellow

Slightly turbid
w/ minute
suspended
particles

9/20/12 6:00
am

Dark Yellow

Clear

9/19/12 8:25
pm

Dark Yellow

Clear

9/20/12 7:45
am

Light Yellow

Clear

9/20/12 10:00
am

Yellow Orange

Turbid

9/20/12 9:55
am

Very light
yellow

Clear

10

9/20/12 9:20
am

Light Yellow

Clear

A. Initial Examination of Urine

Sample
A2. Color

Normal urine is clear and pale yellow

Yellow color is due to the pigment urochrome present
Urine darkens on standing because of oxidation of
urobilinogen to urobilin
Color variations indicate presence of metabolic
abnormality, diseases, stress, ingested foods or drug
Colorless dilution/too much hydration; Diabetes mellitus;
intake of drugs with diuretic effect

Orange

Dark brown, brown red or dark yellow

excessive sweating, concentrated urine

concentrated urine, acute febrile disease

very

Time
collected

Color

Turbidity

pH

9/20/12 7:15
am

Dark yellow

Turbid

9/20/12 7:30
am

Yellow Orange

Clear

9/20/12 9:00
am

Light Pale
Yellow

Slightly Turbid

9/19/12 10:59
pm

Light Yellow

Slightly turbid
w/ minute
suspended
particles

9/20/12 6:00
am

Dark Yellow

Clear

9/19/12 8:25
pm

Dark Yellow

Clear

9/20/12 7:45
am

Light Yellow

Clear

9/20/12 10:00
am

Yellow Orange

Turbid

9/20/12 9:55
am

Very light
yellow

Clear

10

9/20/12 9:20

Light Yellow

Clear

A. Initial Examination of Urine

Sample
A3. Turbidity

Urine may become cloudy due to the presence of

amorphous phosphates which will disappear or due
to urates in urine

Pathologic (e.g. Blood cells, fats, bacteria)

metabolic dysfunction, disease process, or deterioration of
the barrier separating the urinary tract from the blood

Non-pathologic examples are normal crystals,

mucus, epithelial cells, spermatozoa and prostatic fluid ,
cellular casts

Time
collected

Color

Turbidity

pH

9/20/12 7:15
am

Dark yellow

Turbid

9/20/12 7:30
am

Yellow Orange

Clear

9/20/12 9:00
am

Light Pale
Yellow

Slightly Turbid

9/19/12 10:59
pm

Light Yellow

Slightly turbid
w/ minute
suspended
particles

9/20/12 6:00
am

Dark Yellow

Clear

9/19/12 8:25
pm

Dark Yellow

Clear

9/20/12 7:45
am

Light Yellow

Clear

9/20/12 10:00
am

Yellow Orange

Turbid

9/20/12 9:55
am

Very light
yellow

Clear

10

9/20/12 9:20
am

Light Yellow

Clear

A. Gross physicochemical examination of

urine sample
A4. Urine pH

Normal urine pH varies from 4.5 8.0

Urinary pH becomes alkaline because CO2 will diffuse
into the air
The kidney plays a major role in the regulation of
acid-base balance

Reabsorption of sodium and tubular secretion of hydrogen

and ammonium ions

Acidic sodium concentration and excess acid retain by

body

Alkaline bicarbonate-carbonic acid buffer; normally

secreted when there is an excess of base or alkali in the body

Time
collected

Color

Turbidity

pH

9/20/12 7:15
am

Dark yellow

Turbid

9/20/12 7:30
am

Yellow Orange

Clear

9/20/12 9:00
am

Light Pale
Yellow

Slightly Turbid

9/19/12 10:59
pm

Light Yellow

Slightly turbid
w/ minute
suspended
particles

9/20/12 6:00
am

Dark Yellow

Clear

9/19/12 8:25
pm

Dark Yellow

Clear

9/20/12 7:45
am

Light Yellow

Clear

9/20/12 10:00
am

Yellow Orange

Turbid

9/20/12 9:55
am

Very light
yellow

Clear

10

9/20/12 9:20

Light Yellow

Clear

Discussion
B. QUALITATIVE EXAMINATION FOR
NORMAL ORGANIC CONSTITUENTS OF
URINE

TEST FOR UREA

Urea

The end product of protein catabolism

Water-soluble compound
Synthesized exclusively in the liver
Via urea cycle:

involves conversion of ammonia into urea

reactions occur in the liver, but urea is
transported to the kidneys where it is excreted

TEST FOR UREA

TEST FOR UREA

Reagents

Principle

70% NaOH, bromine water

Hydrolyzation of urea with NaOH
Excess NaOH then reacts with bromine water to
form NaOBr, which in turn oxidizes the urea to
form N2,CO2, H2O

Positive result

Evolution of N2 gas as indicated with presence of

bubbles

TEST FOR UREA

MECHANISM

TEST FOR UREA

Group

Test for Urea

White gas was observed then brown gas almost

immediately

Clear yellow solution upper layer and clear light yellow

lower layer

Light yellow solution, with N2

gases, presence of cloudy white substance

Formation of bubbles, light yellow solution

Formation of bubbles on top

Clear yellow with formation of bubbles on the

uppermost portion

Clear yellow solution with presence of bubbles and

foamy layer

Light yellow color upper layer with white bubbles and

clear yellow second layer

Yellow solution with presence of bubbles indicating

formation of N2
gas

10

Evolution of gas was observed

TEST FOR UREA

INDICATIONS
Levels of urea depend upon protein
intake, protein catabolism and kidney
function
Amount of urea excreted per day is 30
g by a normal adult
Elevated urea levels can occur because
of:

Diseases which impair kidney function,

congestive heart failure, diabetes, dietary
changes, liver diseases

TEST FOR URIC ACID

Uric acid
Major end product of catabolism of
purine bases adenine and guanine
nucleotides of cellular DNA and RNA
Formed from dietary nucleic acids
Dissolves in blood and travels to the
kidneys, where it passes out in urine
Excreted in only small amounts in urine
Formula: C H N O
5

TEST FOR URIC ACID

Reagents used

20%Na2CO3, phosphotungstic acid reagent

Principle

Oxidation of alkene in uric acid by

phosphotungstic acid(protein precipitant)
reagent, yielding allantoin and tungsten blue

Positive result

Blue solution

TEST FOR URIC ACID

Reaction Mechanism for the Test for Uric

Acid

TEST For uric acid

Group

Test for Uric Acid

Clear blue solution

Clear light blue solution

A light blue colored solution appeared

Clear blue solution

Formation of blue clear solution

Clear blue solution

Clear light blue solution

Blue solution formed

Light blue solution

10

Light blue colored solution

11

Clear blue solution

TEST FOR URIC ACID

INDICATIONS
High levels of uric acid can cause kidney
stones
Uric acid excretion is 250-750 mg per day
Level of uric acid gradually increases with
age
High levels may be due to:
A high purine diet, gout, and cortisone
therapy.

Gout a painful condition in which high blood

levels of uric acid in the body can cause solid
crystals to form within joints

INDICAN TEST
Indican
An indole produced by bacterial action
on amino acid, tryptophan, in the
intestine
Reflects bacterial activity in the
intestines
Detection in urine depends upon its
decomposition and subsequent
oxidation of indoxyl to indigo blue
Most is excreted in the feces

INDICAN TEST

Reagent

Principle

Obermayers reagent (FeCl3 in

concentrated HCl
and chloroform)
Oxidation of indoxyl to blue indigo and its
absorption into the chloroform layer

Positive result

Blue color of the chloroform layer

INDICAN TEST
MECHANISM

Indican test
Group

Indican Test

Clear blue colored lower layer and dark green upper

layer

Clear brownish green upper layer and blue lower layer

with presence of emulsion

Olive green solution at upper layer and pale clear blue

solution at lower layer

Lower light blue clear solution, middle olive green

clear solution, upper thin black layer

Olive green upper layer and clear light blue lower layer

Clear brown upper layer and very light clear blue lower
layer

Green colored solution upper layer with clear blue lower

layer

No blue color formed at lower layer

Light blue solution at lower layer

10

Light blue colored solution at lower layer

11

Presence of the blue color in the lower layer

INDICAN TEST
INDICATIONS

Increased with high protein diets or

inefficient protein digestion

Excessive urine indicant include:

Maldigestion or malabsorption of
Tryptophan or increased bacterial
overgrowth which reflects enteric toxemia

CREATININE TEST
Creatinine

Anhydride of creatine
A constant constituent of normal urine
Formed from creatine phosphate which is
concerned with muscle contraction
Breakdown of phospocreatine by the addition
of strong acid or alkali. Creatine phosphate
loses phosphoric acid and creatine
undergoes dehydration forming creatinine

CREATININE TEST

Reagents:

Principle:

Alkali picrate solution (5:1, saturated picric

acid and 10% NaOH).
Creatinine reacts with picric acid to form
creatinine picrate complex.

Positive result:

Red-orange colored solution.

CREATININE TEST
MECHANIS
M

CREATININE TEST
Group

Creatinine Test

Clear orange colored solution formed

Clear red orange solution

Dark orange colored solution appear

Red orange colored solution formed

Clear red orange solution formed

Clear red orange solution formed

Clear red orange solution formed

Orange solution formed

Orange solution formed

10

Red colored solution

11

Red orange colored solution

CREATININE TEST
INDICATIONS

All are positive results because creatinine picrate is orange in

color. If the result was red-orange solution, there is a high
presence of creatinine excreted in the urine. However, if the
color is lighter, it means that there is less creatinine in urine
and more in blood because it was not filtered properly.

Decrease of creatinine levels may be due to:

Muscular dystrophy

Increase of creatinine levels may be due to:

Dehydration, bacterial infection, shock, kidney failure,

kidney stones, atherosclerosis

Discussion
C. QUALITATIVE EXAMINATION FOR
PATHOLOGICAL ORGANIC CONSTITUENTS
OF URINE

GUNNINGs TEST
KETONE BODIES

Are products of incomplete fat metabolism

Serve as energy source used in the citric
cycle in place of glucose (when insufficient)
Three water soluble compounds that are
produced as by products when fatty acids are
broken down for energy in the body

GUNNINGs TEST

Test for Ketone bodies

Reagents Used:

Acetone (for positive control only), Conc.

Ammonium hydroxide, Lugols solution

Principle

Halogenation of the enolate of acetone

enolate formation is catalyzed by presence of

hydroxyl ions from NH4OH

Positive result

Iodoform crystals

GUNNINGs TEST
MECHANISM

GUNNINGS
TEST
Group

Urine Sample

Positive Control

Black precipitate settled at

lower layer and cloudy layers
visible on upper layer

Black precipitate was

formed but disappeared.
Turbid solution with cloudy
precipitates at bottom

Dark yellow orange solution

with light yellow precipitate

Dark yellow solution with

light yellow precipitate

Light yellow turbid solution with

black cloud formation seen at
the lower layer/bottom

Pale turbid yellowish solution

with light yellow
precipitate at the bottom;
no black cloud solution
appeared

Clear light yellow solution with

cloudy precipitate

Clear yellow solution with

yellow precipitate

Formation of cloudy white

yellow solution

Formation of white cloudy

precipitate with light yellow
orange solution

GUNNINGS
TEST
Group

Urine Sample

Positive Control

Cloudy yellow solution with

yellow suspension

Clear yellow orange solution

with cloudy white
suspension at the lower part

Turbid/cloudy solution with

precipitate on middle

Slightly turbid/cloudy yellow

solution with black
precipitate at bottom

Clear red orange solution with

cloudy white lower layer and
red orange precipitate at the
bottom

Clear orange solution with

cloudy white lower layer and
yellow orange precipitate
at bottom

Presence of black crystals

settled on the bottom

Presence of black crystals

settled on the bottom

10

Formation of black iodoform

crystals

Turbid solution

11

Yellow solution with cloudy red

precipitate on the bottom

Yellow solution with white

precipitate on the bottom

GUNNINGs TEST
INDICATIONS

Ketonuria ketone bodies present in

urine
Ketosis high level of ketone in blood,
distinct odor
Ketogenesis when liver glycogen is
depleted
Ketoacidosis insulin deficiency, Type 1
diabetes

BENEDICTs test
GLUCOSE

a simple monosaccharide found in plants

serve as an energy source
Usually, there is little or no presence of
glucose in the urine
Glycosuria
Presence of glucose in the urine

BENEDICTs test (Test for Glucose)

Reagents

Principle

Benedicts reagent (carbonate-citrate solution and

copper sulfate solution)
Oxidation reaction of glucose by the Cu2+ and
reduction of Cu2+ to Cu+

Positive Result

Formation of brick red precipitate

BENEDICTs test
MECHANISM

C2. Benedicts Test

Group

Urine Sample

Positive Control

Blue Solution

Yellow-green turbid solution

with yellow precipitate settled
on the bottom

Semi turbid solution with green

precipitate

Turbid orange solution with

orange precipitate

Blue-green colored solution

appeared with formation of a
cloudy-like substance at the
lower part of the test tube

Two layers appeared: a dark

yellow turbid upper layer
solution and a greenish
colored solution for the lower
layer; yellow powdered-like
precipitate formed

Clear blue solution

Upper yellow layer, middle

green layer with orange
precipitate

Pale orange precipitate with

blue-green interphase; yellow
solution

Formation of a clear aqua

solution with suspended
particles

Group

Urine Sample

Positive Control

Clear green upper layer and

clear light blue lower layer with
some particles suspended in
the lower layer and between 2
layers

Orange solution with green

and orange precipitate

Orange solution upper layer;

Green turbid solution middle
layer; Brownish orange
precipitate at lower layer

Slightly clear blue solution

with little precipitate

Blue green turbid upper layer

with few scattered and settling
white precipitate

Turbid yellowish solution

through-out 4 layers with the
third layer being blue green;
Greenish and yellow
precipitate at the bottom

Clear blue solution

Presence of brick red

precipitate

10

Light blue solution

Orange precipitate formed in

an orange solution

Blue solution with a little white

Green turbid solution on top

with orange precipitate on the

11

BENEDICTs test
INDICATIONS:

180mg/dL glucose in urine is the normal

glucose level of a person. 300-500 mg/dL are
common with severe untreated diabetes
Positive result indicates that glucose is
present in the urine. Presence of glucose can
be due to:
Diabetes mellitus; Renal glycosuria

EXTONs TEST
ALBUMIN

Is an umbrella term for a type of protein

which is water soluble
protein is not usually found in urine
Albumin in the human body transports
essential fatty acids from adipose tissue,
otherwise known as fat, to muscle tissue

EXTONs TEST

Test for Albumin

Reagents

Principle

Extons reagent (anhydrous sodium sulfate

and sulfosalicylic acid(SSA) dissolved in
distilled water)
Na2SO4 and SSA will cause acidification and
the application of heat precipitates
albumin

Positive Result

Formation of a white turbid and cloudy

precipitate

Extons Test
Group

Urine Sample

Positive Control

Clear light yellow solution

Creamy white solution with white

precipitate

Turbid light orange solution with fibrous

materials in upper layer and turbid
green with precipitate at lower later

Clear orange solution

Clear light yellow solution

Cloudy pale yellow solution and

formation of light yellow precipitate
at the upper surface

Clear solution

Formation of cloudy mass

suspended in a solution

Formation of clear light yellow

solution

Formation of cloudy solution with

clear bottom

Clear light yellow-orange solution

Light yellow solution with light

yellow aggregates

Clear light yellow solution

White turbid solution with white

precipitate

Clear golden yellow solution

White turbid solution

Clear solution

Cloudiness is seen with mass of

white precipitate

10

Clear colorless solution

White cloudy present

11

No cloudiness appeared

Cloudiness appeared

EXTONs TEST
INDICATIONS

Albuminuria

Presence of albumin in the urine

Proteins from the blood can leak into

the urine when the glomerular filtration
unit of the kidney are
damaged/severed

SMITHs test
BILE PIGMENTS

responsible for lipid catabolism

BILIRUBIN

It is a yellow pigment found in bile, a fluid made

by the liver.
It is secreted in bile and urine.
It is responsible for the brown coloration of feces
and yellow coloration of urine, bruises and
jaundice.
Major constituent of bile.
Conjugated bilirubin is water-soluble and can
therefore be excreted in urine.

SMITHs test

Test for Bile Pigments

Reagents

Principle

Tincture of alcoholic iodine mixture (iodine

crystals and sodium iodide in an absolute
ethanol, then added with distilled water)
Oxidation of the bile pigments to colored
derivatives of the alcoholic iodide

Positive Result

Formation of emerald green at the point of

contact

Smiths Test
Group

Urine Sample

Positive Control

Green interphase

Green interphase

No emerald green interphase

Emerald green interphase

Emerald green interphase

Emerald green interphase

Yellow interphase

Yellow interphase

No emerald green interphase

No emerald green interphase

No emerald green interphase

No emerald green interphase

No emerald green interphase

No emerald green interphase

No emerald green interphase

No emerald green interphase

No emerald green interphase

seen at point of contact

No emerald green interphase at

point of contact

10

No emerald green interphase

Emerald green color appeared

at point of contact

11

Emerald green color is observed

at the point of contact

Emerald green color is

observed at the point of
contact

SMITHs test
INDICATIONS

Dark urine is caused when there is

obstruction to bile flow, or obstructive
jaundice. In which case there is direct
bilirubinemia, which is water soluble and is
filtered through the glomerular membrane.
This is the cause of dark urine.
High levels of bile pigment
(hyperbilirubinemia) in urine may be due to:

Liver injury
Presence of gallstones
Drug toxicity
Hemolytic anemia

Test for occult blood

OCCULT BLOOD

sometimes referred to as hidden blood

Heme groups are most commonly recognized

in their presence as components of
hemoglobin (red pigment in blood)

Can be either hemolyzed (dissolved blood) or

non-hemolyzed (intact red blood cells)

Test for occult blood

Reagents

Principle

spatula of guaiac poweder in 5 mL 95%

ethanol solution; hydrogen peroxide
The peroxidase activity of the Heme which
decomposes H2O2 and the liberated oxygen
oxidizes organic substances such as guaiac
powder

Positive Result

Presence of orange, green ring

Blue ring if much or excess blood is
detected

Test for occult blood

MECHANIS
M

Test for Occult Blood

Group

Urine Sample

Positive Control

Blue ring absent

Blue green ring formed

2
3
4
5
6
7
8
9
10
11

Blue ring absent

Blue ring absent
Blue ring absent
Blue ring absent
Blue ring absent
Blue ring absent
Blue ring absent
Blue ring absent
Blue ring absent
Blue ring absent

Blue green ring formed

Dark blue colored solution appeared
Blue green ring formed
Blue green ring formed
Blue green ring formed
Blue green ring formed
Blue green ring formed
Blue green ring formed
Blue green ring formed
Blue green ring formed

Test for occult blood

INDICATIONS
Hematuria

The presence of more or less intact red

blood cells in the urine
Usually accompanied by hemoglobinuria
because of disintegration of red cell in
urine or brown or produce a smoky
appearance

CONCLUSION

Urine samples from the 11 groups obtained a pH

range of 5-8, most are not turbid and samples in
which suspended particles were seen are
collected for almost 12 hours prior to the
examination that could otherwise allowed the
particles to settled down and provides a false
observation and color of samples varies from
very light yellow to dark orange with respect to
concentration.

Qualitative examination for normal organic

constituents of urine proved that normally it
contains Urea, Uric Acid, Indican and Creatinine.

CONCLUSION

Most urine were found to be negative

for presence of albumin, occult blood,
ketone bodies and bile pigments(with
minor exception for sample nos. 1, 3
and 11.). However 7 out of 11 samples
were proven to be positive for presence
of glucose indicating high sugar
consumption or diet.